ViraPower™ Lentiviral Packaging Mix

인용 및 참조 문헌 (32)

Invitrogen™

ViraPower™ Lentiviral Packaging Mix

ViraPower™ Lentiviral Packaging Mix는 helper 기능을 제공하는 세 가지 packaging plasmid(pLP1, pLP2, pLP⁄VSVG)와 본사 ViraPower™ Lentiviral 발현 시스템을 사용해 lentivirus를자세히 알아보기

카탈로그 번호	수량
K497500	60 reactions

카탈로그 번호 K497500

제품 가격(KRW)

4,346,000

카트에 추가하기

60 reactions

제품 가격(KRW)

4,346,000

카트에 추가하기

ViraPower™ Lentiviral Packaging Mix는 helper 기능을 제공하는 세 가지 packaging plasmid(pLP1, pLP2, pLP⁄VSVG)와 본사 ViraPower™ Lentiviral 발현 시스템을 사용해 lentivirus를 생성하는데 필요한 trans의 구조 및 복제 단백질이 최적화되어 혼합된 mix입니다.

이 제품은 연구용으로만 사용가능합니다. 치료 또는 진단 목적으로 사용할 수 없습니다.

For Research Use Only. Not for use in diagnostic procedures.

구성 또는 유도성 시스템Constitutive

배달 유형Lentiviral

용도(애플리케이션)Viral Expression

제품 유형Lentiviral Packaging Mix

수량60 reactions

선택 제제(진핵)Blasticidin

벡터pLP

클로닝 방법TOPO-TA, Gateway

제품라인ViraPower

프로모터RSV, CMV

단백질 태그V5 Epitope Tag

Unit SizeEach

구성 및 보관

Three tubes, each containing 195 μg lyophilized mix. The packaging mix is enough for 60 transfection reactions in 10 cm tissue culture dishes. Store at -20°C.

자주 묻는 질문(FAQ)

I used one of your lentiviral vectors but am observing cytotoxic effects after transduction. Can you please help?

Possible causes include:

- large volume of viral supernatant used for transduction
- cells sensitive to Polybrene regaent
- too much antibiotic used for selection
- antibiotic used too soon after tranduction
- gene of interest is toxic to cells

I transduced my lentiviral stock into my mammalian cell line but am getting poor expression of my gene of interest. What could have happened?

Poor expression could result from low transduction efficiency, too low of a MOI, too much antibiotic used for selection, usage of antibiotic too soon after transduction, harveting cells too soon after transduction, having a gene of interest that is toxic to cells, or rerrangement in the LTR regions of the expression construct plasmid DNA.

I transduced my lentiviral stock into my mammalian cell line but am getting no expression of my gene of interest. What could have gone wrong?

Here are some possible causes and solutions:
- Promoter silencing; CMV promoter is prone to silencing especially in mouse and rat cells, screen multiple antibiotic resistant clones and select the one with the highest expression levels
- Viral stocks stored incorrectly; aliquot and store at -80 degrees C, do not freeze/thaw more than 3 times

I prepared a lentiviral stock using one of your lentiviral vectors. I am trying to determine the titer using antibiotic selection but am not able to since the cells are very confluent and I am not getting antibiotic-resistant clones. Can you please offer some tips?

Here are some possible causes and solutions:

- Too little antibotic used for selection
- Selection performed on confluent cells; replate cells
- Viral supernatant not diluted sufficiently; titer lentivus using a wider range of 10-fold serial dilutions

I am using one of your lentiviral vectors and am getting a low lentiviral titer. Can you offer some troubleshooting tips?

Possible causes include:

- low transfection efficiency; Use a high-quality plasmid prep, 293FT cells under passage 16, ensure removal of Geneticin during transfection, ensure correct DNA:lipid ratio, and that cells are plated at the correct confluency
- transfected cells are not cultured in medium containing sodium pyruvate; this reagent provides an extra energy source for cells
- viral supernatant harvested too early; viral supernatants can generally be collected 48-72 hrs post-transfection
- viral supernatant too dilute; concentrate virus using CsCl purification
- viral supernatant frozen and thawed multiple times; 3 times should be the maximum freeze/thaw
- gene of interest is large; viral titers decrease as size of insert increases, inserts larger than 5.6 kb are not recommended
- rearrangement in the LTR region of the epxression construct plasmid DNA; use Stb3 cells for transformatin of the lentiviral construct
- poor choice of titering cell line; use HT1080 cells or similar cell line
- Polybrene reagent is not included during transduction; transduce lentiviral construct into cells in the presence of Polybrene reagent
- Lipofectamine reagent handled incorrectly; ensure proper storage and mix gently before use
- Use fluorescence micrscopy to check titer with HiPerform FastTiter lentivirus

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

View all ViraPower™ Lentiviral Packaging Mix FAQs

인용 및 참조 문헌 (32)

인용 및 참조 문헌

Abstract

RIP140-targeted repression of gene expression in adipocytes.

Authors:Christian M, Kiskinis E, Debevec D, Leonardsson G, White R, Parker MG,

Journal:Mol Cell Biol

PubMed ID:16227589

'Ligand-dependent repression of nuclear receptor activity forms a novel mechanism for regulating gene expression. To investigate the intrinsic role of the corepressor RIP140, we have monitored gene expression profiles in cells that express or lack the RIP140 gene and that can be induced to undergo adipogenesis in vitro. In contrast ... More

The membrane anchor R7BP controls the proteolytic stability of the striatal specific RGS protein, RGS9-2.

Authors:Anderson GR, Semenov A, Song JH, Martemyanov KA,

Journal:J Biol Chem

PubMed ID:17158100

'A member of the RGS (regulators of G protein signaling) family, RGS9-2 is a critical regulator of G protein signaling pathways that control locomotion and reward signaling in the brain. RGS9-2 is specifically expressed in striatal neurons where it forms complexes with its newly discovered partner, R7BP (R7 family binding ... More

Constitutively active protein kinase A qualitatively mimics the effects of follicle-stimulating hormone on granulosa cell differentiation.

Authors:Escamilla-Hernandez R, Little-Ihrig L, Orwig KE, Yue J, Chandran U, Zeleznik AJ,

Journal:Mol Endocrinol

PubMed ID:18535249

'Activation of the protein kinase A (PKA) signaling system is necessary for FSH-induced granulosa cell differentiation, but it is not known whether activation of PKA is sufficient to account for the complex pattern of gene expression that occurs during this process. We addressed this question by infecting granulosa cells with ... More

Lysophosphatidic acid-induced expression of periostin in stromal cells: Prognoistic relevance of periostin expression in epithelial ovarian cancer.

Authors:Choi KU, Yun JS, Lee IH, Heo SC, Shin SH, Jeon ES, Choi YJ, Suh DS, Yoon MS, Kim JH,

Journal:Int J Cancer

PubMed ID:20309942

'Lysophosphatidic acid (LPA) is a bioactive lipid crucial for the initiation and progression of ovarian cancer. Identification of LPA-induced biomarkers is necessary for predicting prognosis of ovarian cancer patients. Here we report periostin, an extracellular matrix protein, as an LPA-induced protein in stromal cells and as a prognostic marker in ... More

Id-1 promotes TGF-beta1-induced cell motility through HSP27 activation and disassembly of adherens junction in prostate epithelial cells.

Authors:Di K, Wong YC, Wang X,

Journal:Exp Cell Res

PubMed ID:17916352

'Id-1 (inhibitor of differentiation or DNA binding-1) has been positively associated with cell proliferation, cell cycle progression, and invasiveness during tumorigenesis. In addition, Id-1 has been shown to modulate cellular sensitivity to TGF-beta1 (transforming growth factor beta1). Here we demonstrate a novel role of Id-1 in promoting TGF-beta1-induced cell motility ... More

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